The goal of this Phase I research is to demonstrate the feasibility of developing an anti-sepsis therapeutic. A half million Americans each year are afflicted with sepsis. Of this large number of people, nearly 40 percent or 200,000 people die of this syndrome. Molecular evolution will be used to evolved enhanced properties of acyloxyacyl hydrolase (AOAH), a natural human lysosomal enzyme that detoxifies lipopolysaccharide (LPS or endotoxin), the effector molecule of sepsis. The molecular evolution will increase the affinity of AOAH for LPS to enable it to effectively remove LPS from associating with mediators of sepsis. The major challenge in this investigation is the development of selective procedures that will effectively enrich for AOAH molecules with enhanced properties. Success in Phase I will lead to the isolation of greatly enhanced AOAH anti-sepsis agent in Phase II. Investigations in Phase II will involve in vitro validation of the anti-sepsis character of enhanced AOAH and assessment of the anti-sepsis properties in sepsis animal model systems. PROPOSED COMMERCIAL APPLICATION Sepsis is a major health care problem with half a million people in America contracting this disease each year causing 200,000 fatalities. With increased affinity to LPS (endotoxin) the triggering element in sepsis, acyloxyacyl hydrolase (AOAH), a human enzyme which detoxifies LPS, would have great utility as an anti-sepsis protein therapeutic.